Intranasal Nanodelivery of Oxytocin to Treat Morphine Addiction in HIV Patients by Gene Editing Despite significant developments, treating drug addiction in HIV-1 infected subjects remain as a challenge. Morphine has been shown to exaggerate HIV-induced risk in patients, which deteriorates the brain function and leads to dysregulation of endocrine-metabolic system. This dysregulation might lead to disturbances in the hypothalamic-pituitary-thyroid (HPT) axis, which may indirectly effect the production of Oxytocin (OXT) (neurohypophyseal nona-neuropeptide synthesized in the brain released at the posterior pituitary). Despite the extensive literature on OXT's role in addiction therapy, there are no direct studies available investigating the effects of or on OXT during concurrent Morphine addiction and HIV-1 infection. Also, it has previously been shown that exogenous OXT delivery inhibits the development of acute and chronic morphine tolerance and attenuate the various symptoms of morphine withdrawal in dose dependent manner. OXT may act as a neuromodulator on dopaminergic neurotransmission in limbic-basal forebrain structures to regulate adaptive CNS processes leading to drug addiction. Repeated morphine ingestion will upregulate mu ()-opioid receptor and lead to inhibition of OXT production, which may eventually cause the development of tolerance and physical dependence. Though the OXT has significant therapeutic advantages there are insufficient levels of OXT in compromised diseased state and exogenous OXT expressing source becomes an important need. Exogenous OXT delivery becomes complicated pertaining to factors like plasma half-life and poor oral bioavailability, and limited penetrance of the blood-brain barrier (BBB) due to their large size and hydrophilic nature. Therefore, delivery of a stable and efficacious active gene encoding protein directly into the brain that would reverse the effects of morphine addiction may serve as an effective approach against HIV-1 infected drug abusing subjects. Considering these preclinical limitations related to its delivery and to address this issue, we have developed a novel polyplex nanoformulation of OXT encoding CRISPR activation plasmid by lab developed nontoxic derivative of PEI [P(SiDAAr)5P3] using novel simultaneous spray (SS) technique. Our preliminary results show that SS prepared pGFP polyplex using P(SiDAAr)5P3 (non-viral transfecting agent) could form uniform, small, stable, non-toxic polyplex and achieved significant higher transfection efficiency compared to commercially available jetPEI. To translate this novel technology as anti-addiction therapeutics in HIV-1 treatment paradigm and to achieve the goals, we propose to study the effect of spraying parameters, important formulation aspects and efficacy evaluation of SS-prepared OXT- formulation against Morphine HIV-1 Tat challenge using primary human neurons (Aim-1). generate i nhalable aerosol formulation of OXT- P(SiDAAr)5P3 polyplex Further as a proof of concept, we propose to P(SiDAAr)5P3 polyplex and evaluate intranasal CNS delivery in OXTtm1Wsy/OXTtm1Wsy (OXT deficient- OXTDef) mice exposed to Morphine HIV-1 Tat (Aim-2). Data generated using this grant will be used for future R01 grant proposal.